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GoScript™ Reverse Transcriptase

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Reliable cDNA Synthesis for Sensitive Detection of Expression

  • Available as a standalone enzyme, a complete reverse transcription kit or as a master mix with Oligo(dT) or Random Primers
  • Sensitive transcription of both high-copy and low-copy messages
  • Transcribes short and long transcripts; processes through secondary structure

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Catalog number selected: A5003

$ 172.00
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GoScript™ Reverse Transcriptase
Enzyme/100 reactions
$ 172.00
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Sensitive Reverse Transcriptase Ideal for Challenging Templates

GoScript™ Reverse Transcriptase utilizes M-MLV reverse transcriptase enzyme and state-of-the-art buffer to drive robust, reliable cDNA synthesis of a full range of rare and abundant transcripts, even with difficult templates and in the presence of PCR inhibitors.

Resilient
Sensitive
Reliable
Processive

One Reverse Transcriptase, Four Convenient Formats

GoScript™ Reverse Transcriptase is available in convenient mixes with either Oligo (dT) primers or random primers, as part of a complete kit, and as a stand-alone enzyme.
Complete Kit
Mix + Oligo(dT)
Mix + Random Primers
Standalone Enzyme

GoScript™ Reverse Transcription System

Includes GoScript™ Reverse Transcriptase, Reaction Buffer, MgCl2, dNTP Mix, Oligo(dT), Random Primers and Recombinant RNasin® Ribonuclease Inhibitor, and is designed for efficient synthesis of first-strand cDNA. The system components can effectively reverse transcribe RNA templates starting with total RNA, poly(A)+ mRNA or synthetic transcript RNA.

GoScript™ Reverse Transcription Mix, Oligo(dT)

GoScript™ Reverse Transcription Mix, Oligo(dT) includes an optimized set of reagents designed for efficient synthesis of first-strand cDNA. The Oligo(dT) priming initiates first-strand synthesis by annealing to the 3’ end of any polyadenylated RNA molecule. Reverse Transcription of total RNA or mRNA will yield oligo(dT)-primed products from any poly(A)+RNA. The population of cDNAs produced using this method allows many different downstream analyses from a single reverse transcription reaction.

GoScript™ Reverse Transcription Mix, Random Primers

Includes random primers for synthesis of first-strand cDNA in preparation for qPCR amplification. Priming using random primers is the most generally used method of initiating cDNA synthesis from a variety of RNA templates. Random primers can be used to prime first-strand cDNA synthesis from all RNA molecules, including those that do not possess a poly(A)+ tail and RNA isolated from prokaryotic sources.

GoScript™ Reverse Transcriptase

Includes GoScript™ M-MLV reverse transcriptase enzyme and 5X GoScript™ Reaction Buffer.


Reverse Transcriptase Performance Data

The data presented below illustrate the sensitivity, inhibitor resistance, and performance of GoScript™ Reverse transcriptase in cDNA Synthesis and RT-qPCR.

Efficient cDNA Synthesis Over a Wide Range of Input RNA

efficient cDNA Synthesis Over a Range of Input RNA 13870MB-W

Direct quantification of six different targets in human total RNA using GoTaq® 2-Step RT-qPCR System. Nine parallel large-scale GoScript™ RT reactions were performed, containing log-fold variations of input human reference total RNA (10ng/µl to 0.1fg/µl). 10µl fractions of each reverse transcription reaction product, representing 100ng to 1fg of input RNA, were sampled directly into triplicate GoTaq® qPCR reactions, without dilution, for six separate monoplex assays. Each assay was designed to be specific for a different RNA target with an anticipated, proportionally different expression level: extremely high (5.8S rRNA), high (ACTB and B2M), moderate (HPRT ) or low (RPL29 and TBP1 ). In each different model, the results demonstrate the desired ≅3.3 cycle change per Δlog input, but with different Cqs reflecting relative proportion within the total RNA sample.

Resistance to Common Inhibitors

GoScript reverse transcriptase resistance to inhibitors 8675MB-W

An in vitro-transcribed RNA was reverse transcribed using oligo(dT) primer according to the manufacturer’s instructions. Reverse transcription was carried out in the absence and presence of ethanol (5%, 10% and 20%). cDNA was analyzed by qPCR using GoTaq® qPCR Master Mix. Changes in cycle threshold values in the absence and presence of ethanol were determined and converted to relative change in transcript level = (2Cqinhibitor – Cqnone). Data are the average of three separate experiments.


Efficient Detection of Low-Copy Targets

Here, a series of 10 fold dilutions (1µg - 100pg) of total human RNA (Agilent, Cat #750500) was reverse transcribed using the GoScript™ Reverse Transcription Mix, Random Primers or Oligo(dT).  The resulting cDNA was evaluated by qPCR amplification of the low-copy genes, APC or TBP, using the GoTaq® qPCR Master Mix (Cat. # A6002). Standard curves show that the primers had a calculated efficiency of 99% (APC), and 97% (TBP). Both values are within the range recommended by the MIQE guidelines. No-reverse transcriptase and no-template controls gave negative results.

GoScript™ RT Mix Oligo(dT), TBP Variable RNA Input

GoScript reverse transcriptase  14113MA-W

GoScript™ RT Mix, Random Primers, TBP Variable RNA Input

GoScript reverse transcriptase 14114MA-W

Sensitivity and Specificity

GoScript™ Reverse Transcriptase used in combination with GoTaq® qPCR Master Mix can provide detection of input templates over a 9-log order dynamic range.

Goscript reverse transcriptase dynamic range 8565MB-W-a
Goscript reverse transcriptase titration over 9 logs 8565MB-W-b

GoScript™ Reverse Transcriptase cDNA synthesis coupled with GoTaq® qPCR Master Mix can detect input template over a 9-log order dynamic range. Panel A. Ten RNA template mixes were assembled to model a series of tenfold differences of a specific RNA abundance (1 × 102 to 1 × 1011 copies) in a constant mass of 10ng of human total RNA. cDNAs for each RNA sample were prepared using GoScript™ Reverse Transcriptase and amplified using GoTaq® qPCR at each level of total cDNA (n=3 each point). Inset (Panel A) shows dissociation profile for the 50-, 500- and 5,000-copy standard and 5-copy detected levels of MS2 + cDNA and NTC reactions, demonstrating specificity. Standard curve shown in Panel B illustrates the quantitation and detection of the “unknown” samples. ■ = tenfold standard dilutions; x = detection of 5-copy samples.

Specifications

You are viewing: A5003 Change Configuration

What's in the box?

Item Part # Size Concentration Available Separately

MgCl2

A351H 1 × 1.2ml 25mM View Product

GoScript™ 5X Reaction Buffer

A500C 1 × 600μl

GoScript™ Reverse Transcriptase

A501C 1 × 100μl

Certificate of Analysis

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Use Restrictions

For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.

Storage Conditions

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Specifications

You are viewing: A5004 Change Configuration

What's in the box?

Item Part # Size Concentration Available Separately

MgCl2

A351H 3 × 1.2ml 25mM View Product

GoScript™ 5X Reaction Buffer

A500D 2 × 1.25ml

GoScript™ Reverse Transcriptase

A501D 1 × 500μl

Certificate of Analysis

Search by lot number

Use Restrictions

For Laboratory Use. Outside of the United States, this product is intended for research use only unless otherwise stated.

Storage Conditions

BB

Specifications

You are viewing: A5000 Change Configuration

What's in the box?

Item Part # Size Concentration Available Separately

MgCl2

A351B 1 × 750μl 25mM View Product

GoScript™ 5X Reaction Buffer

A500B 1 × 300μl

GoScript™ Reverse Transcriptase

A501B 1 × 50μl

Oligo(dT)15 Primer

C110B 1 × 50μg 500μg/ml View Product

PCR Nucleotide Mix

C114G 1 × 200μl 10mM View Product

Random Primers

C118B 1 × 50μg View Product

Recombinant RNasin® Ribonuclease Inhibitor

N251A 1 × 2,500u 40u/μl View Product

Nuclease-Free Water

P119A 1 × 1,250μl View Product

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: A5001 Change Configuration

What's in the box?

Item Part # Size Concentration Available Separately

MgCl2

A351H 1 × 1.2ml 25mM View Product

GoScript™ 5X Reaction Buffer

A500C 1 × 600μl

GoScript™ Reverse Transcriptase

A501C 1 × 100μl

Oligo(dT)15 Primer

C110B 1 × 50μg 500μg/ml View Product

PCR Nucleotide Mix

C114G 1 × 200μl 10mM View Product

Random Primers

C118B 1 × 50μg View Product

Recombinant RNasin® Ribonuclease Inhibitor

N251A 1 × 2,500u 40u/μl View Product

Nuclease-Free Water

P119A 2 × 1,250μl View Product

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: A2790 Change Configuration

What's in the box?

Item Part # Size Available Separately

GoScript™ Enzyme Mix

A276A 1 × 100μl

GoScript™ Reaction Buffer, Oligo(dT)

A277A 1 × 200μl

Nuclease-Free Water

P119A 1 × 1,250μl View Product

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: A2791 Change Configuration

What's in the box?

Item Part # Size Available Separately

GoScript™ Enzyme Mix

A276A 2 × 100μl

GoScript™ Reaction Buffer, Oligo(dT)

A277A 2 × 200μl

Nuclease-Free Water

P119A 2 × 1,250μl View Product

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: A2800 Change Configuration

What's in the box?

Item Part # Size Available Separately

GoScript™ Enzyme Mix

A276A 1 × 100μl

GoScript™ Reaction Buffer, Random Primers

A278A 1 × 200μl

Nuclease-Free Water

P119A 1 × 1,250μl View Product

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

Specifications

You are viewing: A2801 Change Configuration

What's in the box?

Item Part # Size Available Separately

GoScript™ Enzyme Mix

A276A 2 × 100μl

GoScript™ Reaction Buffer, Random Primers

A278A 2 × 200μl

Nuclease-Free Water

P119A 2 × 1,250μl View Product

Certificate of Analysis

Search by lot number

Use Restrictions

For Research Use Only. Not for Use in Diagnostic Procedures.

Storage Conditions

BB

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